Cellular and Molecular Physiology, Yale University School of Medicine, New Haven, CT 06511, USA.
Neuron. 2012 Sep 6;75(5):779-85. doi: 10.1016/j.neuron.2012.06.040.
Monitoring neuronal electrical activity using fluorescent protein-based voltage sensors has been limited by small response magnitudes and slow kinetics of existing probes. Here we report the development of a fluorescent protein voltage sensor, named ArcLight, and derivative probes that exhibit large changes in fluorescence intensity in response to voltage changes. ArcLight consists of the voltage-sensing domain of Ciona intestinalis voltage-sensitive phosphatase and super ecliptic pHluorin that carries the point mutation A227D. The fluorescence intensity of ArcLight A242 decreases by 35% in response to a 100 mV depolarization when measured in HEK293 cells, which is more than five times larger than the signals from previously reported fluorescent protein voltage sensors. We show that the combination of signal size and response speed of these new probes allows the reliable detection of single action potentials and excitatory potentials in individual neurons and dendrites.
使用基于荧光蛋白的电压传感器监测神经元电活动受到现有探针响应幅度小和动力学缓慢的限制。在这里，我们报告了一种荧光蛋白电压传感器ArcLight 的开发，以及衍生的探针，这些探针在响应电压变化时表现出荧光强度的大幅变化。ArcLight 由秀丽隐杆线虫电压敏感磷酸酶的电压感应结构域和携带点突变 A227D 的超叶绿素 pHluorin 组成。当在 HEK293 细胞中测量时，ArcLight A242 的荧光强度在 100 mV 的去极化下降低 35%，这比以前报道的荧光蛋白电压传感器的信号大五倍以上。我们表明，这些新探针的信号大小和响应速度的组合允许可靠地检测单个神经元和树突中的单个动作电位和兴奋性电位。
